Taraxacum mongolicum extract induced endoplasmic reticulum stress associated-apoptosis in triple-negative breast cancer cells
Taraxacum mongolicum extract induced endoplasmic reticulum stress associated-apoptosis in triple-negative breast cancer cells
复制标题
DOI：
10.1016/j.jep.2017.04.025
复制
发表时间：
2017-07-12
影响因子：
5.4
通讯作者：
Han, Shu-Yan
中科院分区：
文献类型：
--
作者：
Li, Xiao-Hong;He, Xi-Ran;Han, Shu-Yan
背景：三阴性乳腺癌（TNBC）是一种侵袭性和致命的乳腺癌亚型，治疗选择有限。有必要为跨国公司的管理寻求补充战略。蒲公英（Taraxacum mongolicum，又名蒲公英）是一种具有抗癌活性的中草药，在我国自古以来就被用于治疗乳腺脓肿、乳腺增生等疾病，但其科学依据和作用机制仍有待进一步研究。对蒲公英提取物进行了制备、纯化，并测定了其化学成分。MTT试验计算各组细胞存活率。流式细胞仪检测细胞凋亡和细胞周期。Real-time PCR和Western blotting分别检测mRNA和蛋白质的表达水平。结果：蒲公英提取物能显著降低MDA-MB-231细胞的存活率，诱导细胞发生G2/M期阻滞，并诱导细胞凋亡。同时，它引起裂解的caspase-3和PARP蛋白的显著增加。Caspase抑制剂Z-VAD-FMK可阻断蒲公英提取物诱导的细胞凋亡。蒲公英处理后，三种内质网应激相关信号被强烈诱导，包括ATF 4、ATF 6、XBP 1 s、GRP 78和CHOP基因mRNA表达增加，磷酸化PERK、eIF-2 α、IRE 1以及CHOP和GRP 78下游分子蛋白水平升高。转染CHOP siRNA的MDA-MB-231细胞能显著降低蒲公英提取物诱导的细胞凋亡。结论：蒲公英提取物的抗肿瘤作用可能与ER应激相关的细胞凋亡有关，蒲公英提取物可能是治疗TNBC的有效药物。
Background: Triple-negative breast cancer (TNBC) is an aggressive and deadly breast cancer subtype with limited treatment options. It is necessary to seek complementary strategies for TNBC management. Taraxacum mongolicum, commonly named as dandelion, is a herb medicine with anti-cancer activity and has been utilized to treat mammary abscess, hyperplasia of mammary glands from ancient time in China, but the scientific evidence and action mechanisms still need to be studied.Aim of the study: This study was intended to investigate the therapeutic effect and molecular mechanisms of dandelion extract in TNBC cell line.Methodology: Dandelion extract was prepared and purified, and then its chemical composition was determined. Cell viability was evaluated by MTT assay. Analysis of cell apoptosis and cell cycle was assessed by flow cytometry. The expression levels of mRNA and proteins were determined by real-time PCR and Western blotting, respectively. Caspase inhibitor Z-VAD-FMK and CHOP siRNA were used to confirm the cell apoptosis induced by dandelion extract.Results: Dandelion extract significantly decreased MDA-MB-231cell viability, triggered G2/M phase arrest and cell apoptosis. Concurrently, it caused a markedly increase of cleaved caspase-3 and PARP proteins. Caspase inhibitor Z-VAD-FMK abolished the apoptosis triggered by dandelion extract. The three ER stress-related signals were strongly induced after dandelion treatment, including increased mRNA expressions of ATF4, ATF6, XBP1s, GRP78 and CHOP genes, elevated protein levels of phosphorylated PERK, eIF-2 alpha, IRE1, as well as the downstream molecules of CHOP and GRP78. MDA-MB-231 cells transfected with CHOP siRNA significantly reduced apoptosis induced by dandelion extract. The underlying mechanisms at least partially ascribe to the strong activation of PERK/p-eIF2 alpha/ATF4/CHOP axis.Conclusion: ER stress related cell apoptosis accounted for the anti-cancer effect of dandelion extract, and these findings support dandelion extract might be a potential therapeutic approach to treat TNBC.